Preparation method of chemical luminescence kit for placenta growth factors

A technology of placental growth factor and chemiluminescence reagents, which is applied in the field of preparation of chemiluminescence kits, can solve problems such as detection and identification of trace substances, low cost of excitation solution, low background luminescence, etc. Effect of short, efficient capture, and large specific surface area

Inactive Publication Date: 2019-10-08
NINGBO AUCHEER BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] So far, the methods used to detect placental growth factor residues in human serum mainly include: enzyme-linked immunoassay (ELISA). However, although the detection price of ELISA is low and fast, the sensitivity is not enough, and it is only suitable for the detection of trace substances. Detection and identification; the method adopted in the present invention is the direct chemiluminescence method, and the use of acridinium ester as a chemiluminescent marker has o

Method used

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Embodiment 1

[0022] A preparation method for a chemiluminescence kit of placental growth factor, comprising the following steps:

[0023] (1) Preparation of streptavidin magnetic particle suspension: take streptavidin magnetic particle solution, add TBST solution and mix thoroughly, place on a magnetic separator until the supernatant is free of turbidity, discard the supernatant, and keep Take the magnetic particles, wash them repeatedly 5 times, and dilute them with PBS buffer (50mmol / L, pH=7.8) to obtain a streptavidin magnetic particle solution with a concentration of 0.05%;

[0024] (2) Preparation of biotin-labeled placental growth factor: take 500ug placental growth factor monoclonal antibody, carry out ultrafiltration and centrifugation with a 30KDa ultrafiltration centrifuge tube, and replace PBS buffer (50mmol / L, pH=7.8) with labeling buffer solution, and centrifuged; add biotin stock solution, mix well, and label; after the labeling reaction is completed, add blocking buffer; mL...

Embodiment 2

[0035] A preparation method for a chemiluminescence kit of placental growth factor, comprising the following steps:

[0036](1) Preparation of streptavidin magnetic particle suspension: take streptavidin magnetic particle solution, add TBST solution and mix thoroughly, place on a magnetic separator until the supernatant is free of turbidity, discard the supernatant, and keep Take the magnetic particles, wash them repeatedly 5 times, and dilute them with PBS buffer (50mmol / L, pH=7.8) to obtain a streptavidin magnetic particle solution with a concentration of 0.05%;

[0037] (2) Preparation of biotin-labeled placental growth factor: take 500ug placental growth factor monoclonal antibody, carry out ultrafiltration and centrifugation with a 30KDa ultrafiltration centrifuge tube, and replace PBS buffer (50mmol / L, pH=7.8) with labeling buffer solution, and centrifuged; add biotin stock solution, mix well, and label; after the labeling reaction is completed, add blocking buffer; mL ...

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Abstract

The invention discloses a preparation method of a chemical luminescence kit for placenta growth factors. The preparation method comprises the following steps of preparing streptavidin magnetic particles, and preparing a streptavidin magnetic particle suspension liquid, preparing a placenta growth factor marked by biotin, preparing a placenta growth factor antibody marked by a chemical luminescencesubstance and the like; the streptavidin magnetic particles have high affinity between streptavidin and biotin, so that any biotin-labeled molecules can be bound, and the streptavidin magnetic particles can be used for immunization and molecular detection; the preparation method adopts the polystyrene magnetic particles, and has the characteristics of uniform particle size, large specific surfacearea and regular morphology, so that the target molecule can be rapidly and efficiently captured and magnetic separation is realized; and the magnetic particle chemiluminescence method provided by the invention has strong specificity, accuracy and quickness, short in detection time, accurate in detection result, high in repeatability and the like.

Description

technical field [0001] The invention relates to the field of in vitro diagnosis, in particular to a preparation method of a chemiluminescent kit of placental growth factor. Background technique [0002] Placental growth factor (PLGF) was first isolated and purified from human placental cDNA library by Maglione et al. in 1991. PLGF is mainly synthesized by syncytiotrophoblast cells and can bind to tyrosinase receptors located in trophoblast cells and vascular endothelial cells. It is a protein that has autocrine effects on trophoblast cell functions and paracrine effects on angiogenesis. PLGF has a unique regulatory effect on the function of trophoblast cells and endothelial cells, and can promote angiogenesis. Detecting the blood PLGF level of pregnant women can be used clinically to identify the oxygen supply pressure of placental syncytiotrophoblast cells, and to predict, identify and monitor treatment of preeclampsia. [0003] So far, the methods used to detect placenta...

Claims

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Application Information

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IPC IPC(8): G01N33/74G01N33/543G01N33/532G01N21/76
CPCG01N33/74G01N33/54326G01N33/532G01N21/76
Inventor 周义正唐静黄丹娣
Owner NINGBO AUCHEER BIOTECHNOLOGY CO LTD
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